JP2009011260A - Tray-formed container and method for filling material to be housed into the same container - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a tray-formed container, giving a uniform cell density per unit based on the cell culturing region of the bottom surface of the container, also without concentrating the cells at the peripheral edge of the container or a part of the same, while suppressing the travel of the cells and suitable for multiplying the cells. <P>SOLUTION: This tray-formed container is provided with a container main body 1 obtained by opposing a film-state first container wall 1a and second container wall 1b, hermetically sealing a peripheral edge 1c, and at least one of ports 1f, 1g installed at the container main body 1; wherein a multiple number of recessed parts 10, 11... are installed on the first container wall 1a, for suppressing the travel of the housed material to the extended presenting direction of the first container wall 1a, and also a multiple number of housing chambers are installed for uniformly subdividing the housed material with the recessed parts 10, 11... of the first container wall 1a and the second container wall 1b. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a tray-like container suitable for culturing animal cells such as lymphocytes collected from a patient in a closed system aseptically and safely and efficiently, and a method for filling the container with the contents. is there.

  In recent years, adoptive immunotherapy has been performed in which cells collected from patients are cultured outside the body and the proliferated cells are returned to the body. For culturing and proliferating cells, a flask or petri dish, and a bag-like container having a sealed inner periphery and a housing space inside are used.

  In culturing cells using the container described above, select a container of a size that matches the number of cells and the volume of the medium (hereinafter referred to as “container”), add the culture solution in a timely manner, It is necessary to transfer the contained contents to a container of a different size according to the capacity. This work is not only complicated, but there is a risk of causing contamination such as microorganisms such as bacteria, mycoplasma and viruses.

  In addition, if the container is placed at an angle, or if it is left unattended, distortion of the film in the bag-like container, or slight vibration from the surrounding environment or incubator, etc. may cause the cells in the container to be contained in the container. The concentration of the cells in the vicinity and the vicinity of the periphery, and the damage caused by the physical collision of the cells and the cell density in the liquid phase are increased, thereby inducing the proliferation of the cells.

  In cell culture using a bag-like container, oxygen necessary for cell growth is ensured through the film constituting the bag. The number of cells per area is extremely large, oxygen necessary for cell proliferation cannot be secured, and cells are excessively concentrated to induce apoptosis and cell death, resulting in a decrease in cell proliferation rate.

  In addition to the container described above, a well plastic plate is used in which a substantially rectangular parallelepiped container body having a plurality of recesses provided on the bottom surface is covered with a lid. Has to be injected or recovered for each of the recesses, requiring laborious work. In addition, since the injection or recovery operation is performed with the lid removed, there is a risk of causing contamination such as microorganisms such as bacteria and mycoplasmas and viruses.

Furthermore, focusing on well-structured cell culture containers, conventionally, a flask-type container has been proposed in which the container body has a substantially rectangular parallelepiped shape and a plurality of U-shaped recesses are provided on one surface of the container body. This culture vessel has been proposed for suspension culture of cells capable of forming aggregates such as neural stem cells. (Patent Document 1)
JP 2006-055069 A

  This container has a reduced diameter structure in which the bottom area is smaller than the opening area of the recess to intentionally form an aggregate of cells regardless of the cell density. It is difficult to secure oxygen necessary for the cells to be grown and the cells on the bottom side of the flask to grow. Further, as with a general flask, there is a possibility that contamination such as microorganisms such as bacteria and mycoplasma and viruses may occur in the injection or recovery operation.

  The present invention, for example, when used as a cell culture container, in the cell culture region on the bottom surface of the container, by suppressing the movement of cells in the extending direction of the bottom surface of the container, the cells are concentrated on the periphery or part of the container. An object of the present invention is to provide a tray-like container suitable for growing cells without doing so.

  The present invention eliminates the time-consuming work of injecting or recovering the contained material for each recess, which is necessary when using a well-structured plastic container having a plurality of recesses on the bottom surface, and bacteria. An object of the present invention is to provide a tray-like container that can reduce contamination of microorganisms, mycoplasmas, viruses, and the like.

Claim 1 of the present application is a tray having a container body whose peripheral edge is sealed with the first container wall and the second container wall facing each other, and at least one port communicating with a space in the container body. In the container,
The first container wall is provided with a plurality of recesses for suppressing the contents in the container body from moving in the extending direction of the first container wall.

  In the present invention, the port includes not only a short tubular body having a shape to which other connecting members can be easily connected, but also a soft tube integrated with the container body by fusion or the like. It is a small-diameter opening having a configuration capable of filling the container body with the contained material and discharging the contained material from the container body.

  In addition to the case where all the contents in the container main body are divided into a plurality of recesses and accommodated, when the volume of the contents exceeds the total volume of the plurality of recesses, a part of the contents is a recess. However, this is also included in the present invention.

  A second aspect of the present invention is the tray-shaped container according to the first aspect, wherein the concave portion has a flat bottom surface.

  A third aspect of the present invention is the tray-shaped container according to the first or second aspect, wherein the concave portions have substantially the same bottom area and depth.

  According to a fourth aspect of the present invention, in the tray-like container according to any one of the first to third aspects, the second container wall is planar and can contact the top of the partition wall that defines the recess. It is characterized by that.

  Claim 5 of the present application is the tray-like container according to any one of claims 1 to 3, wherein the second container wall is planar, and is in a non-contact state with the top of the partition wall that partitions the recess. It is characterized by maintaining.

  A sixth aspect of the present invention is the tray-like container according to any one of the first to third aspects, wherein the second container wall has a convex portion that can be inserted into the concave portion.

  A seventh aspect of the present invention is the tray-like container according to any one of the first to sixth aspects, wherein the top of the partition wall that defines the concave portion has a concave portion that can communicate with the adjacent concave portion. To do.

  An eighth aspect of the present invention is the tray-like container according to any one of the first to seventh aspects, wherein at least one of the first container wall and the second container wall has oxygen permeability.

  A ninth aspect of the present invention is the tray-shaped container according to any one of the first to eighth aspects, wherein the inside of the container body can be seen through at least one of the first container wall and the second container wall. And

A tenth aspect of the present invention is the tray-like container according to any one of the first to ninth aspects, wherein the concave portion has a depth of 0.7 mm or more and 20 mm or less.
In addition, as for the depth of the said recessed part, 2 mm or more and 15 mm or less are more preferable.

The eleventh aspect of the present invention is the tray-shaped container according to any one of the first to tenth aspects, wherein the bottom surface of the concave portion has an area of 0.3 cm ² or more and 20 cm ² or less.

  Claim 12 of the present application is the tray-like container according to any one of claims 1 to 11, wherein the contents in the container body are injected or embedded in an animal, or cells to be attached to an injured part. It is characterized by including.

Claim 13 of the present application is a container main body having a film-shaped first container wall and a second container wall facing each other and having a peripheral edge sealed,
At least one port communicating with the space in the container body;
A plurality of recesses that are provided on the first container wall and prevent the contents in the container body from moving in the extending direction of the first container wall;
A method for filling things in a tray-like container comprising:
A step of accommodating the container in the container body from the port;
Dividing the contents in the container body into the plurality of recesses by the weight of the contents and / or the pressing force in the thickness direction applied to the container body;
And clogging the port.

Claim 14 of the present application is a tray having a container main body whose peripheral edge is sealed with the first and second container walls facing each other, and at least one port communicating with the space in the container main body. In the container,
Provided in the container body is a partition wall that forms a plurality of compartments that abut against the first container wall and inhibit the contents in the container body from moving in the extending direction of the first container wall. It is characterized by that.

Hereinafter, the case where the effect of the present invention is used as a cell culture container will be described as an example.
According to the tray-like container according to claim 1 of the present application, the first container wall is provided with a plurality of recesses for suppressing the contents in the container body from moving in the extending direction of the first container wall. The cells in the medium in the container body are divided into a plurality of recesses, settled to the bottom surface of each recess, and the movement of the first container wall in the extending direction is suppressed. For this reason, when the tray-like container is placed at a slight inclination, or even if the first container wall is distorted, the cells in the medium do not concentrate on the peripheral edge or part of the container body, It becomes possible to proliferate cells without causing a decrease in the proliferation rate of the cells due to apoptosis.

  According to the tray-like container of claim 1 of the present application, the periphery of the container body having a plurality of recesses is sealed, and at least one port communicating with the space in the container body is provided. Therefore, the medium containing cells can be filled into the container body from the port, and the medium containing cells can be easily divided into a plurality of recesses by, for example, slightly tilting the tray-like container in the length direction or the width direction. .

  Further, according to the tray-like container according to claim 1 of the present application, at the time of recovering the proliferated cells, for example, by inverting and tilting the tray-like container, it can be easily discharged from the port. As in the case of using a plastic container, it is possible to eliminate the time-consuming work of injecting or collecting cells in each recess using a pipette. Furthermore, since the cells can be filled and the proliferated cells can be recovered while maintaining the closed system, contamination such as microorganisms such as bacteria and mycoplasmas and viruses can be reduced.

  According to the tray-like container according to claim 2 of the present application, since the recess has a flat bottom surface, it is possible to prevent the cells in the medium from being concentrated in one place even in each recess.

  According to the tray-like container according to claim 3 of the present application, the bottom area and the depth of each of the concave portions are substantially equal, so that the cell density can be made substantially uniform for each concave portion.

  According to the tray-like container according to claim 4 of the present application, the second container wall is flat and can contact the top of the partition wall that divides the recess, so that the second container wall is on the first container wall side. The medium containing the cells can be easily divided into the respective recesses by pressing to.

  According to the tray-like container according to claim 5 of the present application, the second container wall is planar and maintains a non-contact state with the top of the partition wall that divides the recess, so that the volume of the medium containing cells is plural. It is suitable for the case where the total volume of the recess is exceeded.

  According to the tray-like container according to claim 6 of the present application, since the second container wall has a convex part that can be recessed into the concave part, even if the medium containing cells does not reach the entire volume of the plurality of concave parts. The medium containing cells can be divided into substantially uniform amounts in the respective recesses.

  According to the tray-like container according to claim 7 of the present application, by having a recessed portion that can communicate with the adjacent recessed portion at the top of the partition wall that partitions the recessed portion, a medium containing cells that are divided into the recessed portions, Since it can be made to flow into an adjacent recessed part via a recessed part, the culture medium containing a cell can be disperse | distributed to each recessed part substantially equally easily.

  According to the tray-like container according to claim 8 of the present application, since at least one of the first container wall and the second container wall has oxygen permeability, it is possible to reliably ensure oxygen necessary for cell growth.

  According to the tray-like container according to claim 9 of the present application, since the inside of the container body can be seen through from at least one of the first container wall or the second container wall, without taking out the cells in the container body at any time, Cells can be observed visually or with a microscope from the outside of the container body.

  According to the tray-shaped container according to claim 10 of the present application, since the concave portion has a depth of 0.7 mm or more and 20 mm or less, it is possible to suppress the movement of the settled cells in a stationary state, and the first container wall Uneven thickness when forming the recess can be reduced. In addition, it is more preferable that the concave portion has a depth of 2 mm or more and 15 mm or less, and by setting the concave portion to 2 mm or more, the sedimented cells can be removed from the concave portion even when the container is moved or vibrations occur during the culture. In addition, by setting the thickness to 15 mm or less, uneven thickness generated at the time of forming the concave portion can be suppressed within an allowable range, and stable molding can be performed.

According to the tray-like container of claim 11 of the present application, since the bottom surface of the recess has an area of 0.3 cm ² or more and 20 cm ² or less, it is possible to suppress the induction of apoptosis due to excessive concentration of cells.

  According to the tray-like container according to claim 12 of the present application, since the contents in the container main body contain cells to be injected or embedded in the animal, or stuck to the damaged part, In addition, the risk of causing contamination such as microorganisms such as bacteria and mycoplasmas and viruses can be reduced, and cells that are injected or embedded in animals or that are attached to damaged parts should be more suitably cultured. Can do.

According to the method of filling the contents into the tray-like container according to claim 13 of the present application, the process of accommodating the contents in the container body from the port;
A step of dividing the contents in the container body into a plurality of recesses by the weight of the contents and / or the pressing force in the thickness direction applied to the container body;
Clogging the port;
Therefore, the time-consuming work of injecting cells into each recess, such as when using a plastic container having a well structure, can be omitted.

  According to the tray-like container according to claim 14 of the present application, the container main body is brought into contact with the first container wall, and the contents in the container main body are prevented from moving in the extending direction of the first container wall. Since the partition which forms a some division is provided, the effect substantially the same as the tray-shaped container which concerns on Claim 1 of this application can be achieved.

  The illustrated embodiment exemplifies a case where the tray-like container according to the present invention is used as a cell culture container, and will be described below based on the illustrated embodiment. Of course, the tray-like container according to the present invention can be used not only for cell culture containers but also for other purposes.

  As shown in FIGS. 1 and 2, the tray-like container according to the present embodiment is a container body in which the peripheral edge 1c is sealed with the flexible film-like first container wall 1a and second container wall 1b facing each other. 1 and two ports 1 f and 1 g welded to the container body 1. In addition, the container body 1 is provided with suspension holes 1d and 1e that are suspended as bags on the left and right sides on the upper end side.

  First, the ports 1f and 1g welded to the container body 1 will be described. The ports 1f and 1g are flow paths for injecting cells, a medium, and the like into the container body 1, and the port 1f is a cell or medium. A port 1g is provided for injecting a medium or the like and connecting a filling tube 21 for connection with another culture solution container. The port 1g is used for sampling the contents in the container body 1. It is provided to connect port 3.

  Each of the ports 1f and 1g may be a tubular molded product formed from a thermoplastic resin such as polyethylene, polypropylene, vinyl chloride, or a tube. Preferably, a container formed of the same material as the container wall 1a or 1b is provided.

  The filling tube 21 includes a connection port 20 branched into two, and can be fitted to the port 1 f or 1 g of the container body 1. One tube 21d of the filling tube 21 is provided with an inlet 21a for injecting a culture solution or the like into the container body 1, and the inlet 21a is provided with an inlet cap 21b and an inlet cover 21c. ing. The other tube 22d is provided with a connection port 22a for connection to another bag filled with a culture solution, and the connection port 22a is provided with a connection port cap 22b and a connection port cover 22c.

  Here, the injection port 21a and the connection port 22a are not particularly limited as long as there is an opening for taking in and out the contents such as culture medium and culture medium containing cells, but those having a lure shape are more preferable. More preferred are those used in infusions, blood bags and the like.

  The sampling port 3 is a sealing plug having a connection port 3a for connecting to the port 1f or 1g and a rubber plug 3c for piercing the injection needle, and the injection needle piercing portion of the rubber plug 3c is sealed. Covered with a stop cap 3b.

  The first container wall 1a is formed by additionally processing a film-like member in a concavo-convex shape. The first container wall 1a has a container such as a medium containing cells in the container body 1, particularly a container. When the main body 1 is kept horizontal, the concave portions 10, 11... For suppressing the cells that have settled on the inner surface of the first container wall 1a from moving in the extending direction of the first container wall 1a are A plurality (49 in FIG. 1) are provided in the width direction. Moreover, the bottom area and depth of each recessed part 10,11 ... are substantially the same. 3 to 5, reference numeral 10 a indicates the bottom surface of the recess 10.

  A plurality of storage chambers (same as “recesses”) that can divide the contents are formed from the recesses 10, 11... Of the first container wall 1a and the second container wall 1b. 1b is formed of a planar film material covering the recesses 10, 11,... Of the first container wall 1b.

  Examples of film materials constituting the first container wall 1a and the second container wall 1b include low density polyethylene, linear low density polyethylene, polypropylene, ethylene vinyl acetate copolymer, cyclic polyolefin, methylpentane resin, polybutadiene resin, and styrene. Examples thereof include those made of a thermoplastic resin such as butadiene styrene block copolymer and vinyl chloride. This film is not limited to a single layer, but may be a multi-layered film in which the inner surface side is made of a soft resin as a sealant layer and the outer surface side is made of a crystallized resin.

  At least one of the first container wall 1a and the second container wall 1b is formed of an oxygen permeable film. For example, the second container wall 1b is formed of an oxygen permeable film, and the first container wall 1a is formed of either an oxygen permeable film or a non-permeable film.

The oxygen permeable film referred to here is oxygen in a gas environment in an environment of 25 ° C., and the oxygen permeation amount is 1000 cc / m when tested by a test method in accordance with JIS K 7126 “Gas permeability test of plastic film and sheet” It refers to not more ² / 24h / atm or more, oxygen permeability is 2000cc ^/ m 2 ^/ 24h ^/ atm or higher is more preferable. Thereby, the oxygen required for cell growth can be ensured.

  In addition, at least one of the first container wall and the second container wall is formed of a transparent or translucent film that can be seen through. Thereby, without collecting the cells in the container main body 1, it is possible to observe the growth state of the cells and the like from the outside of the container main body 1 visually or with a microscope at any time.

  When the recesses 10, 11,... Of the first container wall 1a have a depth of 0.2 mm or more and 20 mm or less, preferably 0.7 mm or more and 20 mm or less, the cells that have settled in a stationary state move. Can be suppressed, and uneven thickness when the concave portion of the first container wall is formed can be reduced. In addition, it is more preferable that the concave portion has a depth of 2 mm or more and 15 mm or less, and by setting the concave portion to 2 mm or more, the sedimented cells can be removed from the concave portion even when the container is moved or vibrations occur during the culture. In addition, by setting the thickness to 15 mm or less, uneven thickness generated at the time of forming the concave portion can be suppressed within an allowable range, and stable molding can be performed.

Further, by forming the bottom surfaces of the recesses 10, 11,... In a planar shape and having an area of 0.3 cm ² or more and 20 cm ² or less, induction of apoptosis caused by excessive concentration of cells can be suppressed. Here, in order to supply oxygen necessary for proliferation to each cell and to prevent the occurrence of apoptosis caused by excessive concentration of cells, the volume of one recess 10 is 0.5 to 20 cc. About 1 to 8 cc is more preferable.

  In the illustrated embodiment, the bottom surface of the recesses 10, 11,... Of the first container wall 1a is formed to have a square shape in plan view, but the shape of the bottom surface of the recesses 10, 11,. The shape may be a rhombus, hexagon, octagon, circle, or the like.

  As shown in FIG. 3, the second container wall 1b is composed of a flat film surface covering the recesses 10 of the first container wall 1a, and adjacent recesses 10 and 11 provided on the first container wall 1a. It is configured to be able to contact the top surface 10c of the partition wall 10b formed therebetween. Thereby, since the culture medium containing the cell divided into the accommodation chamber can be covered with a flat film surface, the cell density per unit area can be kept even during the culture.

  In addition, although the 2nd container wall 1b is comprised with a planar film surface, you may comprise so that the 2nd container wall 1b may not contact the top surface 10c of the division wall 10b.

The embodiment shown in the drawings will be described more specifically below. The first container wall 1a is formed of a polyethylene film having a thickness of 350 μm. The first container wall 1a provided with the recesses 10, 11,... Is an uneven shape formed by vacuum forming with a mold provided with 49 rectangular parallelepipeds having a bottom area of 1.44 cm ² and a height of 7 mm. It is formed using the film which has the shape of.

The second container wall 1b is an oxygen permeation amount ^{2000cc / m 2 / 24h / atm} , which is formed by using a polyethylene film of 100μ thickness, thereby, become oxygen necessary for cell culture and can be secured ing.

  In the tray-shaped container configured as described above, since the medium containing cells is divided and stored in the recesses (accommodating chambers) 10, 11..., The settled cells concentrate on a part of the container body 1, the periphery, and the like. And the cell density per unit area can be kept substantially uniform.

  For this reason, oxygen necessary for proliferation can be supplied to each cell, and the occurrence of apoptosis caused by excessive concentration of cells can be prevented, and the cell growth rate can be reduced without causing a decrease in cell proliferation rate. Proliferation can be continued. Moreover, since it is a closed system as a container, the risk of contamination such as microorganisms such as bacteria and mycoplasmas and viruses can be reduced.

  In the process of proliferating cells, the distance between the cells is important, but since the distance between the cells suitable for cell proliferation can be maintained, the proliferation can be enhanced by stimulating the cells. This is thought to prevent apoptosis caused by cell density due to cell concentration being too high, and avoiding cell proliferation signal transmission and cell growth not proceeding at low cell density. Furthermore, since the cell density can be adjusted, it is possible to culture with the optimal amount of medium and cytokines. Not only can the loss be lost, but the cell density can be increased within the range of cell proliferation to increase the amount of culture medium and cytokines used. And the size of the container can be reduced, so that a space saving effect can be obtained.

  Furthermore, although the tray-like container of the present embodiment has a three-dimensional structure, since the basic material is made of a film, it is easily crushed unlike a hard container that is generally used. Can also be obtained.

  Moreover, since the periphery of the 1st container wall 1a and the 2nd container wall 1b is sealed, a tray-like container is a closed system and systemization of a closed system becomes easy. In addition, since the container is a closed system, the risk of contamination such as microbes such as bacteria and mycoplasmas and viruses can be reduced, and cells injected into or embedded in animals or attached to damaged areas can be reduced. It is suitable as a container used for culture.

  In the above-described embodiment, the second container wall 1b has been described based on the case of forming a flat film surface. However, as shown in FIG. 4, the second container wall 1b is a recess 10 of the first container wall 1a. It can form so that it may have the convex part 12 which can be indented. Thereby, when the culture medium containing the cells in the container main body 1 is less than the entire volume of the recesses 10, 11,... Of the first container wall 1a, the recesses 10, 11,. The volume can be reduced, and even when the medium containing the cells in the container body 1 is relatively small, the medium can be divided into the recesses 10, 11,.

  In addition, as shown in FIG. 5, the concave portions 10 of the first container wall 1a can be formed to have concave portions 13 communicating with adjacent concave portions on the top surface 1c of the partition wall 1b. As a result, the medium containing the cells in each storage chamber can be easily separated and dispersed by flowing into the adjacent recesses by the recess 13.

  In relation to the above-described oxygen permeability, in general, a method of forming a film thickness as thin as 100 to 150 μm is employed to provide oxygen permeability. For example, the first container wall 1a is formed using this thin film. 6 and 7, it is possible to hold the first container wall 1a in a three-dimensional shape by accommodating the first container wall 1a in a separately formed holding tray 4 as shown in FIGS. It becomes.

  The holding tray 4 has a flange portion 4a on which the peripheral edge 1c of the container body 1 can be placed, and is formed to have a peripheral side wall 1b and a bottom surface 1c that can accommodate the recesses 10 of the first container wall 1a. . Further, the holding tray 4 can be formed as a plurality of concave portions corresponding to the respective concave portions 10 of the first container wall 1a.

  Furthermore, in order to ensure oxygen permeability, at least one vent hole 4d is provided on the bottom surface 1c, or after at least one vent hole 4d is provided on the bottom surface 1c, a fine mesh sheet is placed on the bottom surface of the holding tray 4. And the bottom surface of the recess 10... So that the recess 10... Of the first container wall 1 a can be kept flat and oxygen can permeate into the container body 1 from the entire recess of the first container wall. It is.

  Next, using the tray-like container, the contents (for example, a medium containing cells) can be filled into the container body 1 by the following procedure. First, as shown in FIG. 8, the injection port cover 21c and the injection cap 21b are removed from the injection port 21a of one tube 21d of the filling tube 21, and the outer cylinder 5 of the syringe is inserted into the injection port 21a. Is injected into the container body 1.

  After injecting the contents, as shown in FIG. 9, after removing the outer cylinder 5 of the syringe from the injection port 21a, the container body 1 is kept in the lying state. Thereafter, the container main body 1 is shaken, or the container main body 1 is inclined in the length direction or the width direction, and the contents are distributed to be divided into a plurality of concave portions 10, 11. Then, the second container wall 1b is pressed in the thickness direction to discharge the internal air of the container body 1 from the injection port 21a, and finally the injection port 21a is sealed with the injection cap 21b to form a closed bag. .

  By using this tray-like container, a time-consuming work such as injecting or recovering the contents of each recess using a pipette like a well-structured plastic container having a plurality of recesses on the bottom surface. It can be omitted. Moreover, since it can be used as a closed system container, the risk of causing contamination such as microorganisms such as bacteria and mycoplasmas and viruses can be reduced.

  In order to confirm the effectiveness of the tray-like container of the present invention, the following experiment was conducted using a tray-shaped container having a plurality of concave portions and a flat bottom surface as a comparative example together with the tray-shaped container of the present invention. .

RPMI medium (containing IL-2) containing 5% of autologous serum (1.5 × 10 ⁷ CELLS) collected from a healthy person: The contents added to 50 mL of the container body 1 from the injection port 21a After injecting into the interior, the injection cap 21b was closed, and the contents were slowly mixed while keeping the container body 1 horizontal.

  Next, after removing the injection cap 21b again, the air inside the container body 1 is taken out while slowly pressing the second container wall in the thickness direction, thereby removing the recesses 10, 11,. The container is accommodated in each of the recesses 10, 11... In a substantially uniform manner close to the top surface 1 c of the partition wall 1 b to be partitioned, so that the cell density in each of the recesses 10, 11. Was covered with a second container wall.

On the other hand, RPMI medium containing 5% of autologous serum of activated lymphocytes (1.5 × 10 ⁷ CELLS) collected from a healthy person, as usual, in a tray-like container without a plurality of recesses and having a flat bottom surface (Contains IL-2): Cultured cells added to 50 mL were filled.

  Each container was placed in an incubator, and the cultured cells were cultured at 37 ° C. and saturated humidity in a 5% carbon dioxide atmosphere for 3 days. Next, 50 mL of the culture solution RPMI was added to the inside of each container that had been cultured for the previous 3 days to make the capacity in each container 100 cc, and then placed in the incubator again and cultured for 3 days.

As a result, the final number of cells in each container and the number of cells per 1 cm ^{2 of} culture area were 3.0 × 10 ⁸ (5.1 × 10 ⁶ per 1 cm ^{2 of} culture area) CELLS in the tray-like container of the present invention. . In a tray-like container having a plurality of concave portions and having a flat bottom surface as a comparative example, it was 1.9 × 10 ⁸ (3.2 × 10 ⁶ per 1 cm ^{2 of} culture area) CELLS.

  In the above-described embodiment, the first container wall 1a is provided with the recesses 10, 11,..., But the first container wall 1a and the second container wall 1b are formed in a planar shape, and the first container wall In the container main body 1 formed between 1a and the second container wall 1b, the inner surface of the first container wall 1a is brought into contact with the contents in the container main body 1 in the extending direction of the first container wall 1a. A plurality of depressions are formed by arranging a plurality of partition walls that suppress movement, for example, by arranging a lattice-shaped partition wall, using the lattice-shaped partition wall as a peripheral surface and the first container wall 1a as a bottom surface. You may do it.

It is a top view which shows embodiment of the tray-shaped container which concerns on this invention. It is a side view which shows the tray-shaped container of FIG. It is a partial sectional side view which shows the tray-shaped container of FIG. It is a fragmentary sectional side view which shows another embodiment with the tray-shaped container of FIG. It is a fragmentary perspective view which shows embodiment which provided the recessed part in the partition wall of the tray-shaped container of FIG. It is a top view which shows embodiment which provided the holding tray in the tray-shaped container of FIG. It is a fragmentary sectional side view which shows embodiment which provided the holding tray in the tray-shaped container of FIG. It is explanatory drawing which shows the filling process of the accommodation thing by the tray-shaped container of FIG. It is explanatory drawing which shows the accommodation process following the filling process of the stored matter by the tray-shaped container of FIG.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Container body 1a 1st container wall 10,11 Recessed part (accommodating chamber)
1b Second container wall 1f, 1g Port

Claims (14)

In a tray-like container comprising a container body having a film-shaped first container wall and a second container wall facing each other and having a peripheral edge sealed, and at least one port communicating with a space in the container body,
A tray-like container comprising a plurality of recesses in the first container wall for suppressing the contents in the container body from moving in the extending direction of the first container wall.   The tray-like container according to claim 1, wherein the recess has a flat bottom surface.   The tray-like container according to claim 1 or 2, wherein the concave portions have substantially the same bottom area and depth.   The tray-like container according to any one of claims 1 to 3, wherein the second container wall has a planar shape and can contact a top of a partition wall that partitions the recess.   The tray-like container according to any one of claims 1 to 3, wherein the second container wall is planar and maintains a non-contact state with a top of a partition wall that partitions the recess.   The tray-like container according to any one of claims 1 to 3, wherein the second container wall has a convex part that can be recessed into the concave part.   The tray-like container according to any one of claims 1 to 6, further comprising a recessed portion that can communicate with an adjacent recessed portion at a top of a partition wall that partitions the recessed portion.   The tray-like container according to any one of claims 1 to 7, wherein at least one of the first container wall and the second container wall has oxygen permeability.   The tray-like container according to any one of claims 1 to 8, wherein the inside of the container main body can be seen through from at least one of the first container wall or the second container wall.   The tray-like container according to any one of claims 1 to 9, wherein the concave portion has a depth of 0.7 mm or more and 20 mm or less. The tray-like container according to any one of claims 1 to 10, wherein a bottom surface of the concave portion has an area of 0.3 cm ² or more and 20 cm ² or less.   The tray-like container according to any one of claims 1 to 11, wherein the contents in the container main body include cells to be injected or embedded in an animal or attached to a damaged part. A container body having a film-like first container wall and a second container wall facing each other and hermetically sealed at the periphery;
At least one port communicating with the space in the container body;
A plurality of recesses that are provided on the first container wall and prevent the contents in the container body from moving in the extending direction of the first container wall;
A method for filling things in a tray-like container comprising:
A step of accommodating the container in the container body from the port;
Dividing the contents in the container body into the plurality of recesses by the weight of the contents and / or the pressing force in the thickness direction applied to the container body;
Clogging the port;
A method for filling a container into a tray-like container, comprising: In a tray-like container comprising a container body having a film-shaped first container wall and a second container wall facing each other and having a peripheral edge sealed, and at least one port communicating with a space in the container body,
Provided in the container body is a partition wall that forms a plurality of compartments that abut against the first container wall and inhibit the contents in the container body from moving in the extending direction of the first container wall. A tray-like container characterized by that.

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